1. Field of the Invention
The present invention relates to antiviral compositions based upon constituents isolated from or contained within medicinal mushroom mycelia, or the corresponding synthetic molecules, that are shown to be useful in reducing pathogenic viruses, and treating viral infections; in particular viruses that afflict animals, including, but not limited to, humans, bees, pigs, bats, and birds, resulting in a reduction of disease causing viruses, their pathogenicity and/or infectivity in both the animal host and the environment.
2. Description of Related Art Including Information Disclosed Under 37 CFR 1.97 and 1.98
Medicinal mushrooms have been used for thousands of years for a wide assortment of ailments. Traditionally the mushroom fruitbody has been used. Scientists have extensively studied extracts of the fruitbodies over the past decades. Although numerous papers have been published showing hot water extracts of mushrooms and their mycelia can activate immune systems and can be anti-inflammatory, comparatively few have elucidated the benefits of the alcohol fractions. The current invention describes novel contributions to the field of medicinal mushroom research, particularly discoveries pertaining to antiviral activity of alcohol extracted mushroom mycelium and the active constituents contained within them.
Scientists are now discovering that viral infection challenges and degrades the immune system in multiple ways including inflammation, which can lead to cellular damage from free radicals, a cofactor in carcinogenesis, and to cancers caused by oncoviruses. Worldwide, the World Health Organization (WHO) International Agency for Research on Cancer estimated that in 2002 17.8% of human cancers were caused by infection, with 11.9% being caused by 1 of 7 different viruses. The 7 viruses that are known to cause cancer include three herpes oncoviruses: Epstein-Barr aka human herpesvirus 4 (HHV-4); human herpesvirus 6 (HHV-6) and human herpesvirus 8 (HHV-8). HHV-6 is implicated in the development of lymphomas, leukemia, cervical cancers, Karposi sarcoma, and brain tumors. Other oncoviruses include the polyoma virus that causes Merkel cell carcinoma (MCC), the human papillomaviruses (HPV 16 and 18) which cause cervical cancer, anal cancer, oropharyngeal cancers, vaginal cancers, vulvar cancers and penile cancers; hepatitis B and C, which cause liver cancer; and the human T-lymphotropic viruses (HTLV), which cause T-cell leukemia and T-cell lymphoma. Four HTLVs are well known. HTLV-1 and HTLV-2 are involved in epidemics, infecting 15-20 million people worldwide. In the United States, hepatitis C infection is estimated at 2.7 million and 700,000-1.4 million persons are estimated to be infected with hepatitis B. HTLVs can be more prevalent in some geographical regions than others, infecting around 1% of Japan's population. Rates among volunteer blood donors in the U.S. average 0.016% but in parts of Africa, reports of 15% have been recorded.1 
As science progresses, more oncoviruses and virally-mediated oncogenic pathways are likely to be discovered. Since immunity is based on many complex factors, pathogenic viruses which have not been known to specifically cause cancer may contribute to carcinogenesis by causing inflammation, free radicals, and reducing the number of immune cells that would otherwise keep cancer and co-infections at bay. Nature is a number's game, and the balance between health and disease is imperiled by infections. When immunity is lowered, the human body is less able to eradicate cancer cells, which would otherwise be kept in check, and deleterious inflammatory pathways further challenge health. Thus mortality rates rise with compounded infections. Hence there is a need for compositions that reduce oncoviruses and also reduce viruses that cause inflammation and immune deactivation, contributory to oncogenesis.
Viral epidemics and pandemics represent increasing threats to global health as zoonotic diseases spread, jumping host species, recombining, and potentially mutating into more virulent forms. The need for additional anti-influenza drugs is important in maintaining active countermeasures against pandemics. As an example, when H1N1 flu virus swept the world in 2010, the two most popularly effective antivirals were Tamiflu® (Oseltamivir) and Relenza® (Zanamivir); which were useful, at best, for shortening the disease period by approximately a day. In less than one year, the novel H1N1 virus evolved to increasingly resist Tamiflu® applications, and the drug has largely become ineffective against downstream populations of the heritage H1N1 swine flu viruses. Although some antiviral medicines may be effective at present, it is vital that researchers investigate diverse therapeutic agents in order to combat viral outbreaks from rapidly evolving strains. The ease and speed with which numerous varieties of flu virus mutate to become drug resistant is particularly concerning.
In the spring of 2015, wild birds from Asia carried H5N8 viruses to North America, which co-mingled with bird flu variants and mutated into a highly pathogenic H5N2 virus. This virus resulted in the killing—both from the virus and euthanasia—of tens of millions of birds and threatened the multibillion dollar chicken and turkey industry. The H5N2 virus has mutated into H5N1 variants, and given the number of hosts in wild and domesticated birds, continued mutations could evolve a strain of the flu that could leap to humans, causing a pandemic and severe devastation to our global economies, our food biosecurity and human health. Given that flu viruses can be spread via airborne, direct and secondary contacts (via vehicles, shoes, clothing, washcloths, dollar bills, flies, mites, etc.) and that flu viruses can survive in mucous droplets for up to 17 days, the threat of a flu pandemic spreading to humans greatly concerns specialists in virology, public health and defense. Finding methods and compositions to reduce the viral pathogen payloads vectored by host animals and fomites will greatly serve the public interest. Moreover, since most vaccines have limited (but focused) utility against only a few flu variants, finding broad based solutions to preventing and reducing the threat from multiple flu viruses in particular, and diverse viruses in general, is of paramount importance.
Medicinal mushrooms have been ingested as food and as therapy for hundreds, and in some cases, thousands of years. This is strong support for their safe ingestion, making them appealing candidates in the search for new antiviral agents. In addition, the compounds disclosed herein may be resident ingredients within well-known foods, which, when isolated and concentrated can function as drugs. The difference here then between a food and a drug is that a drug is typically an isolated molecule presented in a form at a high purity (i.e. >90%), and used at a high dose in treating a disease. One of the first mushrooms recognized for its antiviral activity was Fomes fomentarius, a hoof-shaped wood conk that was found to inhibit the tobacco mosaic virus.2 More recently, derivatives of the Gypsy mushroom, Rozites caperata, were found by Piraino et al. to significantly inhibit the replication and spread of varicella zoster (the ‘shingles’ virus), influenza A and B, and herpes simplex I and II. Sarkar et al. have also identified activity against herpes simplex I in an extract of Shiitake mushrooms (Lentinula edodes).3,4 Collins and Ng have identified a polysaccharopeptide inhibiting HIV type 1 infection from Turkey Tail mushrooms (Coriolus versicolor=Trametes versicolor).5 Brandt and Piraino, and Stamets have also published summaries of the antiviral properties of some mushrooms species.6,7 
The prevailing adamant belief by those skilled in the science of medicinal mushroom research is that the only benefits from medicinal mushroom extracts must come from hot water extraction. As three noted experts, skilled in the art, and authors of scientific papers and books on the medicinal properties of mushrooms, have published: “Hot water extracts are the only form of mushroom preparation ever used in Traditional Chinese Medicine (TCM), and the only form of mushroom supplement ever used, tested or studied in the scientific and medical research.” (The Health Benefits of Medicinal Mushrooms, 2005, Dr. Mark Stengler).8 
According to John Seleen of Mushroom Science (currently on his Mushroom Science website), “Few people realize how much research has been conducted on medicinal mushrooms; more than 2,000 studies have been published in just the last 10 years, and all of these studies have used hot water extracts. In fact, hot water extracts are the only type of medicinal mushroom preparation that has actual proof of effectiveness for supporting immune health . . . . It is not often that you have absolute consensus between 1,000's of years of herbal practice and every scientific study ever published on that same subject, but that is the case with medicinal mushrooms. All sources and traditions agree, medicinal mushrooms must be extracted with hot water when used for immune support, and hot water extracts are the only type of mushroom supplement validated by the research.” (Sep. 10, 2015).
Additionally, a 2015 ‘white paper’ by Jeffery S. Chilton, Redefining Medicinal Mushrooms: A new scientific screening program for active compounds states that if mycelium is grown on rice, and not wood, that “Without the natural precursors, basidiomycete mycelium in sterile culture produces few of the important secondary metabolites.” Thus these three experts, skilled in the art, and greatly influential, are unanimous in discrediting any significant activity of non-hot water mycelial extracts, especially when mycelium is grown on grains such as rice. Thus they teach away from the specifics of this invention.
With the advent of tissue culture of mycelium in the early part of the 20th century, this new mushroom life stage (the mycelium as opposed to the mushroom fruit bodies) became available for testing bioactivity. This newly available fungal form opened up new frontiers for natural product research. However, pharmaceutical companies studying mushroom-based natural products, typically and more inexpensively, analyze the fruitbodies, and in doing so miss the antiviral activities this inventor has discovered that are expressed during the mycelium life stage.
From a practical point of view, pharmaceutical researchers find it easier to collect and analyze a mushroom rather than to laboriously culture it and then analyze the mycelium. This standard approach has a reasonable rationale: many species of mushroom forming fungi do not grow, or are too slow to grow in in vitro culture compared to other fungi such as molds. Additionally, the mushroom fruitbodies are made of compacted mycelium—dense with tissue—and hence would seemingly be a better resource for bioprospecting than the more loosely netted mycelium. This would explain why there is little prior art on the mycelium of mushroom species being anti-virally active. Typically, when a pharmaceutical company screens mushroom-based natural products, they analyze large sets of species. If they do not find activity in the natural form (the mushroom fruitbody or carpophore), they move on to other species without further exploring a negative result, based upon the mistaken belief that the activities of the mushrooms would be the same as the mycelium and that all strains or cultivars of a species would possess the same antiviral activity. This is understandable since the prevailing belief is that the mushroom is simply composed of compacted mycelium and the two would share, in common, the same constituents.
Recent genomic research shows that more genes are turned on during the mycelial stage of development than in the reproductive structure of the mushroom fruitbodies. As was noted by Li et al., 2013, “The protein-coding genes were expressed higher in mycelia or primordial stages compared with those in the fruiting bodies.”9 The inventor's practices have inadvertently laid claim to or supported this statement without prior knowledge that more genes are up-regulated during mycelial growth than fruitbody (mushroom) formation. This was not known, nor obvious, at the time when this patent applicant filed his first provisional antiviral patent application U.S. 60/534,776 on Jan. 6, 2004.
Remarkably, and unexpectedly, the author's discovery that the alcohol soluble extracts of Ganoderma lucidum (Ganoderma lucidum var. resinaceum) mycelium showed anti-flu activity is novel in that it is in direct contradiction to past results that alcohol extracts from fruitbody extracts had no activity. This is unique in that it is counter-intuitive as conventional thinking would lead most scientists to believe that the activity of both forms would share commonality of effects.
Seong-Kug Eo tested both water soluble and alcohol soluble fractions from the fruitbodies of Ganoderma lucidum.10 The methanol soluble compounds were labeled as “GLMe,” for “Ganoderma lucidum methanol fraction” and “GLhw” for “G. lucidum hot water.”—Their conclusions showed that the methanol (alcohol) soluble fractions had no activity against flu viruses: “The carpophores of G. lucidum (500 g) were disrupted and extracted with hot water for 8 h. The water extract was concentrated to a 10th of the original volume, and three volumes of ice cold EtOH added to precipitate the high molecular weight components. After standing out overnight at 4° C., it was centrifuged and the precipitates were lyophilized, and GLhw (3.30 g) as a brownish substance was obtained. Eight methanol soluble substances (GLMe) were isolated by organic solvents on the basis of differences in the net electric charge. GLMe-1, -2, -4 and -7 isolated from the MeOH fraction exhibited inhibitory effects, especially on the cytopathic effects induced by VSV Indiana and New Jersey strains at concentrations which did not show cytotoxicity against Vero cells; however, they exhibited no effect on the other viruses such as HSV and influenza A virus.”
The author notes that this article has been referenced, as of this date, Aug. 18, 2015, 2,090 times according to Google and 3,570 times by Bing search engines, showing that Seong-Kug Eo et al.'s (1999) statement that the alcohol soluble fractions of Ganoderma lucidum were inactive against flu and herpes viruses was well established in the scientific literature.
This discovery by Seong-Kug Eo et al. (1999) teaches away from Stamets U.S. Pat. No. 8,765,138 (2014), the latter of which discloses that alcohol extracts of the Ganoderma lucidum mycelium were highly active against flu and herpes viruses. Conventional wisdom has been that the fruitbodies (mushrooms) of Ganoderma lucidum held the most diverse bioactive constituents and that the mycelium is less active. Moreover, since the fruitbodies are composed of mycelium, that there would be differences between extracts made from mushrooms vs mycelia would have been seen, by those currently and historically skilled in preparations of mushrooms, to be a factual contradiction. Hence, the inventor's results were both novel and nonobvious at the time of this inventor's first antiviral patent applications wherein he found that the EtOH/H2O extracts of fruitbodies of Agarikon (Fomitopsis officinalis) were inactive against pox, flu and herpes viruses whilst the EtOH/H2O extracts from the living mycelium of this species were active against the same viruses.